The objectives of this proposal are: (1) To develop a productive cultural medium of catharanthus roseus cell culture for production of antineoplastic indole alkaloids. (2) To develop an immobilized cell (or enzyme) system and a bioreactor-extractor for biosynthesis of the desired alkaloids. Addition of certain metal ions has been found to induce accumulation of new alkaloids in the cell culture and the proximal precursors (catharanthine and vindoline) for the antineoplastic alkaloids. This opens up new ways to elucidate the biosynthetic pathway. Intermediates normally undetectable due to fast metabolism may be accumulated and identified. Chemical bioregulators (known to have effect in vivo) will be synthesized and added to induce alkaloid production in cell culture. A bioreactor-extractor using immobilized cells/enzymes is designed to synthesize continuously the end product alkaloids with the least supplementation of cofactors.